Osteocytes control myeloid cell proliferation and differentiation through GSα-dependent and -independent mechanisms

INTRODUCTION: Previous studies have shown that osteocytes, the matrix-embedded cells in bone, control bone modeling and remodeling through direct contact with adjacent cells and via secreted factors that can reach cells in the bone marrow microenvironment (BMM). Osteocytes express several receptors including G protein-coupled receptors (GPCRs) and mice lacking the stimulatory subunit of G-proteins (Gsα) in osteocytes have abnormal myelopoiesis, skeletal abnormalities and reduced adipose tissue. This study aimed at evaluating the effects of osteocyte-secreted factors on myeloid cell proliferation and differentiation in vitro. To investigate cross-talk between osteocytes and the BMM, we established osteocytic cell lines lacking Gsα expression to study the molecular mechanisms by which osteocytes control myeloid cell proliferation and differentiation. METHODS: CRISPR/Cas9 was used to knockout Gsα in the osteocytic cell line Ocy454. Conditioned media (CM) from differentiated Ocy-GsαCtrl and Ocy-GsαKO cells were used to treat myeloid cells and bone marrow mononuclear cells (BMNCs) isolated from long bones of 6-8-week-old C57/BL6 mice. BMNCs were cultured with Macrophage Colony Stimulating Factor (M-CSF), Receptor Activator of Nuclear Factor Kappa β Ligand (RANKL) to induce osteoclast differentiation. Proliferation, TRAP staining, TRAP activity, resorption pit assay, F-actin ring formation and mRNA expression were used to evaluate cell proliferation, differentiation and function of the induced osteoclasts. Proteomics analysis of CM was performed to identify osteocyte-secreted factors capable of controlling myelopoiesis and osteoclastogenesis. RESULTS: Myeloid cells treated with CM from Ocy-GsαKO showed a significant increase in cell proliferation compared to Ocy-GsαCtrl CM and non-treated control. BMNCs treated with CM from Ocy-GsαCtrl and Ocy-GsαKO showed a significant increase in cell proliferation as compared to non-treated control. Osteoclast differentiation was significantly suppressed by CM from Ocy-GsαCtrl and further suppressed by CM from Ocy-GsαKO compared to non-treated control. Osteoclasts exposed to CM from Ocy-GsαKO showed a significant defect in activity and function as compared to cells exposed to CM from Ocy-GsαCtrl and non-treated cells. Osteoclast apoptosis was significantly enhanced by Ocy-GsαCtrl and Ocy-GsαKO CM compared to non-treated control. Among osteocyte secreted factors, we identified neuropilin-1 (NRP-1) as a Gsα-dependent osteocytic factor capable of suppressing osteoclastogenesis. CM from Ocy-GsαKO in which M-CSF was reduced by shRNA demonstrated decrease in BMNCs proliferation, demonstrating that osteocytes are also important sources of this cytokine. CONCLUSIONS: Osteocytes produce several Gsα-dependent and -independent secreted factors capable of supporting myelopoiesis, promoting macrophage proliferation and suppressing osteoclast formation. We identified osteocyte-derived NRP-1 as a novel factor capable of decreasing osteoclastogenesis. In addition, we found that M-CSF secreted by osteocytes is responsible in part for BMNC proliferation. Future studies should focus on determining the role of osteocyte-mediated NRP-1 and other secreted factor(s) in control of myelopoiesis and osteoclastogenesis.2020-06-26T00:00:00

Outdoor thermal comfort optimization through vegetation parameterization : species and tree layout

The optimization of outdoor thermal comfort has become the keystone to guarantee the healthy and comfortable use of outdoor spaces. This study aims to optimize the outdoor thermal comfort through vegetation parameterization in a boulevard located in Guelma city, Algeria during summertime. However, two main parameters were investigated, species and tree layout, through a numerical simulation. We first collected microclimate data of a sunny summer day. Second, we used real microclimate data in different simulations using the Envi-met atmospheric model. The findings reveal that Ficus Nitida is the most significant species to intercept solar radiation and provide shade over the day in Souidani Boudjemaa Boulevard, with a maximum reduction of Ta = 0.3 °C and UTCI = 2.6 °C at 13:00 p.m. Tree layout is a determining parameter in the creation of shaded paths, based on the quality of the shadows cast by the trees, namely, their size. Thereby, planting the washingtonia palm trees along the center of the boulevard is the best option to maximize the shaded area within the boulevard, with maximum reduction of Ta = 1.8 °C and UTCI = 3.5 °C at 16:00 p.m.https://www.mdpi.com/journal/sustainabilityMechanical and Aeronautical Engineerin

Reducing the environmental impact of surgery on a global scale: systematic review and co-prioritization with healthcare workers in 132 countries

Abstract Background Healthcare cannot achieve net-zero carbon without addressing operating theatres. The aim of this study was to prioritize feasible interventions to reduce the environmental impact of operating theatres. Methods This study adopted a four-phase Delphi consensus co-prioritization methodology. In phase 1, a systematic review of published interventions and global consultation of perioperative healthcare professionals were used to longlist interventions. In phase 2, iterative thematic analysis consolidated comparable interventions into a shortlist. In phase 3, the shortlist was co-prioritized based on patient and clinician views on acceptability, feasibility, and safety. In phase 4, ranked lists of interventions were presented by their relevance to high-income countries and low–middle-income countries. Results In phase 1, 43 interventions were identified, which had low uptake in practice according to 3042 professionals globally. In phase 2, a shortlist of 15 intervention domains was generated. In phase 3, interventions were deemed acceptable for more than 90 per cent of patients except for reducing general anaesthesia (84 per cent) and re-sterilization of ‘single-use’ consumables (86 per cent). In phase 4, the top three shortlisted interventions for high-income countries were: introducing recycling; reducing use of anaesthetic gases; and appropriate clinical waste processing. In phase 4, the top three shortlisted interventions for low–middle-income countries were: introducing reusable surgical devices; reducing use of consumables; and reducing the use of general anaesthesia. Conclusion This is a step toward environmentally sustainable operating environments with actionable interventions applicable to both high– and low–middle–income countries

The utility of the interferon gamma-inducible protein-10 (IP-10) level in bronchoalveolar lavage and blood in the diagnosis of tuberculosis

Introduction: Control of TB depends on early detection and treatment of active cases. Aim of the work: Investigating the utility, sensitivity and specificity of interferon gamma inducible protein IP-10 in both blood and bronchoalveolar lavage (BAL) in the diagnosis of TB infection in clinically suspected patients. Methods: Thirty patients with clinical and/or radiological suspicion of pulmonary tuberculosis and negative sputum smear for AFB with Z–N stain were included in the study. BAL and blood samples were sent for the estimation of the level of interferon gamma inducible protein IP-10. Results: IP-10 levels in both blood and BAL were significantly higher in TB patients (P = 0.005 and 0.007 respectively). Sensitivity of IP-10 in blood was 100% and specificity was 60%. Positive predictive value was 56%, negative predictive value was 100% and accuracy was 73%. Sensitivity of IP-10 in BAL was 100% and specificity was 35%. Positive predictive value was 44%, negative predictive value was 100% and accuracy was 57%. Sensitivity of IP-10 in blood and BAL were similar (100%) and both were more sensitive than tuberculin skin test (TST) (sensitivity 67%). Detection of IP-10 in blood (specificity = 60%) was more specific than its detection in BAL (specificity = 35%). On the other hand, specificity of detection of IP-10 in blood (60%) was comparable to the tuberculin test (specificity = 62%). Conclusion: Interferon gamma inducible protein IP-10 may help in detecting M. tuberculosis infection and monitoring disease activity and efficacy of therapy

Combined Zircon/Apatite U-Pb and Fission-Track Dating by LA-ICP-MS and Its Geological Applications: An Example from the Egyptian Younger Granites

Laser Ablation-Inductively Coupled Plasma-Mass Spectrometry (LA-ICP-MS) is classically used in U-Pb dating to measure U and Pb isotopic concentrations. Recently, it has become frequently used in fission-track (FT) chronometry too. As an advantage, the U-Pb and FT double dating will enable efficiently determining the crystallization ages and the thermo-tectonic history concurrently as samples volume, analytical time, efforts, and cost will be greatly reduced. To demonstrate the validity of this approach, a Younger granite (Ediacaran age) sample from North Eastern Desert (NED), Egypt was analyzed for U-Pb and FT double dating. The integration of multiple geochronologic data yielded a zircon U-Pb crystallization age of 599 ± 30 Ma, after emplacement, the rock cooled /uplifted rapidly to depths of 9–14 km as response to the post-Pan African Orogeny erosional event as indicated by apatite U-Pb age of 474 ± 9 Ma. Afterwards, the area experienced a slow cooling/exhumation for a short period, most-likely as response to denudation effect. During the Devonian, the area was rapidly exhumed to reach depths of 1.5–3 km as response to the Hercynian tectonic event, as indicated by a zircon FT age of 347 ± 16 Ma. Then the studied sample has experienced a relatively long period of thermal stability between the Carboniferous and the Eocene. During the Oligocene-Miocene, the Gulf of Suez opening event affected the area by crustal uplift to its current elevation. This integration of Orogenic and thermo-tectonic information reveals the validity, efficiency, and importance of double dating of U-Pb and FT techniques using LA-ICP-MS methodology

Boosting of Antioxidants and Alkaloids in Catharanthus roseus Suspension Cultures Using Silver Nanoparticles with Expression of CrMPK3 and STR Genes

Global agricultural systems are under unprecedented pressures due to climate change. Advanced nano-engineering can help increase crop yields while ensuring sustainability. Nanotechnology improves agricultural productivity by boosting input efficiency and reducing waste. Alkaloids as one of the numerous secondary metabolites that serve variety of cellular functions essential for physiological processes. This study tests the competence of silver nanoparticles (AgNPs) in boosting alkaloids accumulation in Catharanthus roseus suspension cultures in relation to the expression of C. roseus Mitogen Activated Protein Kinase 3 (CrMPK3) and Strictosidine Synthase (STR) genes. Five concentrations (5, 10, 15, 20 and 25 mg·L−1) of AgNPs were utilized in addition to deionized water as control. Results reflected binary positive correlations among AgNPs concentration, oxidative stress indicated with increase in hydrogen peroxide and malondialdehyde contents, activities of ascorbate peroxidase and superoxide dismutase, expression of the regulatory gene CrMPK3 and the alkaloid biosynthetic gene STR as well as alkaloids accumulation. These correlations add to the growing evidence that AgNPs can trigger the accumulation of alkaloids in plant cells through a signaling pathway that involves hydrogen peroxide and MAPKs, leading to up-regulation of the biosynthetic genes, including STR gene

Defens in gene expression in some plant sources of Taif

Plant defensins are promising future strategy as antimicrobial agents; however the number of characterized plant defensins is very low. We isolated and studied the expression of plant defensins in six plants from Taif region. Using RT-PCR and two pairs of common as well as three pairs of specific primers, the defensin gene expression was analyzed. Plants showed differences in defensin gene expression in floral buds and leaves, though floral buds represented higher gene expression. The amino acid sequence of the six isolated cDNA sequences showed high similarity with other defensin accessions from the nucleotide database, especially in the cysteine rich motif. Phylogenetic analysis revealed that the isolated sequences share the common features of plant defensins, especially the 8 conserved cysteines. The results of this study help to detect some valuable Saudi plant sources for the development of natural peptides as a replacement for chemical antibiotics

Green Synthesis of Zinc Oxide Nanoparticles (ZnO-NPs) Using Arthrospira platensis (Class: Cyanophyceae) and Evaluation of their Biomedical Activities

In this study, zinc oxide nanoparticles (ZnO-NPs) were successfully fabricated through the harnessing of metabolites present in the cell filtrate of a newly isolated and identified microalga Arthrospira platensis (Class: Cyanophyceae). The formed ZnO-NPs were characterized by UV–Vis spectroscopy, Fourier transform infrared (FT-IR), transmission electron microscopy (TEM), energy-dispersive spectroscopy (EDX), X-ray diffraction (XRD), and X-ray photoelectron spectroscopy (XPS). Data showed the efficacy of cyanobacterial metabolites in fabricating spherical, crystallographic ZnO-NPs with a size ≈30.0 to 55.0 nm at a wavelength of 370 nm. Moreover, FT-IR analysis showed varied absorption peaks related to nanoparticle formation. XPS analysis confirms the presence of Zn(II)O at different varied bending energies. Data analyses exhibit that the activities of biosynthesized ZnO-NPs were dose-dependent. Their application as an antimicrobial agent was examined and formed clear zones, 24.1 ± 0.3, 21.1 ± 0.06, 19.1 ± 0.3, 19.9 ± 0.1, and 21.6 ± 0.6 mm, at 200 ppm against Bacillus subtilis, Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli, and Candida albicans, respectively, and these activities were reduced as the NPs concentration decreased. The minimum inhibitory concentration (MIC) values were determined as 50 ppm for S. aureus, 25 ppm for P. aeruginosa, and 12.5 ppm for B. subtilis, E. coli, and C. albicans. More interestingly, ZnO-NPs exhibit high in vitro cytotoxic efficacy against cancerous (Caco-2) (IC50 = 9.95 ppm) as compared with normal (WI38) cell line (IC50 = 53.34 ppm)